dc.date.accessioned | 2020-12-01T20:23:13Z | |
dc.date.available | 2020-12-01T20:23:13Z | |
dc.date.created | 2020-11-24T15:49:02Z | |
dc.date.issued | 2021 | |
dc.identifier.citation | Aeschimann, Walter Kammer, Stephan Staats, Stefanie Schneider, Petra Schneider, Gisbert Rimbach, Gerald Cascella, Michele Stocker, Achim . Engineering of a functional γ-tocopherol transfer protein. Redox Biology. 2021, 38 | |
dc.identifier.uri | http://hdl.handle.net/10852/81326 | |
dc.description.abstract | α-tocopherol transfer protein (TTP) was previously reported to self-aggregate into 24-meric spheres (α-TTPS) and to possess transcytotic potency across mono-layers of human umbilical vein endothelial cells (HUVECs). In this work, we describe the characterisation of a functional TTP variant with its vitamer selectivity shifted towards γ-tocopherol. The shift was obtained by introducing an alanine to leucine substitution into the substrate-binding pocket at position 156 through site directed mutagenesis. We report here the X-ray crystal structure of the γ-tocopherol specific particle (γ-TTPS) at 2.24 Å resolution. γ-TTPS features full functionality compared to its α-tocopherol specific parent including self-aggregation potency and transcytotic activity in trans-well experiments using primary HUVEC cells. The impact of the A156L mutation on TTP function is quantified in vitro by measuring the affinity towards γ-tocopherol through micro-differential scanning calorimetry and by determining its ligand-transfer activity. Finally, cell culture experiments using adherently grown HUVEC cells indicate that the protomers of γ-TTP, in contrast to α-TTP, do not counteract cytokine-mediated inflammation at a transcriptional level. Our results suggest that the A156L substitution in TTP is fully functional and has the potential to pave the way for further experiments towards the understanding of α-tocopherol homeostasis in humans. | |
dc.language | EN | |
dc.rights | Attribution-NonCommercial-NoDerivatives 4.0 International | |
dc.rights.uri | https://creativecommons.org/licenses/by-nc-nd/4.0/ | |
dc.title | Engineering of a functional γ-tocopherol transfer protein | |
dc.type | Journal article | |
dc.creator.author | Aeschimann, Walter | |
dc.creator.author | Kammer, Stephan | |
dc.creator.author | Staats, Stefanie | |
dc.creator.author | Schneider, Petra | |
dc.creator.author | Schneider, Gisbert | |
dc.creator.author | Rimbach, Gerald | |
dc.creator.author | Cascella, Michele | |
dc.creator.author | Stocker, Achim | |
cristin.unitcode | 185,15,12,59 | |
cristin.unitname | Teoretisk kjemi | |
cristin.ispublished | true | |
cristin.fulltext | postprint | |
cristin.qualitycode | 1 | |
dc.identifier.cristin | 1851816 | |
dc.identifier.bibliographiccitation | info:ofi/fmt:kev:mtx:ctx&ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=Redox Biology&rft.volume=38&rft.spage=&rft.date=2021 | |
dc.identifier.jtitle | Redox Biology | |
dc.identifier.volume | 38 | |
dc.identifier.doi | https://doi.org/10.1016/j.redox.2020.101773 | |
dc.identifier.urn | URN:NBN:no-84407 | |
dc.type.document | Tidsskriftartikkel | |
dc.type.peerreviewed | Peer reviewed | |
dc.source.issn | 2213-2317 | |
dc.identifier.fulltext | Fulltext https://www.duo.uio.no/bitstream/handle/10852/81326/5/1-s2.0-S2213231720309782-main.pdf | |
dc.type.version | PublishedVersion | |
cristin.articleid | 101773 | |
dc.relation.project | NFR/262695 | |